US4594328A — Tissue protein PP21, a process for obtaining it a…

Claim 1

Independent Claim 170 words

A protein PP 21 , extracted from a source of said protein and having the following characteristics: (a) an electrophoretic mobility in the region of that of β 1 -globulins; (b) an isoelectric point of 4.6±0.3; (c) a sedimentation coefficient s 20 ,w of 3.2±0.2 S; (d) a molecular weight determined in an ultracentrifuge of 52,900±6,200; (e) an extinction coefficient E 1cm 1% (280 nm) of 10.5±1.0; (f) a carbohydrate fraction of 19.2±5.2 g/100 g (mannose 1.8±0.4, galactose 4.3±1.0, fucose 1.3±0.3, N-acetylglucosamine 6.5±2.0 and N-acetylneuraminic acid 5.3±1.5, each g/100 g); and (g) an amino acid composition as in the following table: ______________________________________ Coefficient Amino acid Residues per 100 residues of variation ______________________________________ lysine 8.75 3.91 histidine 1.58 6.81 arginine 2.75 6.07 aspartic acid 9.59 1.97 threonine 3.73 7.22 serine 7.43 5.95 glutamic acid 8.16 5.15 proline 11.34 9.36 glycine 7.01 1.25 alanine 4.98 3.62 cystine 1/2 5.50 0.84 valine 8.14 1.42 methionine 0.00 0.00 isoleucine 4.86 2.78 leucine 5.42 10.34 tyrosine 5.01 5.01 phenylalanine 3.56 1.00 tryptophan 2.12 17.65 ______________________________________

Claim 2

Dependent Claim 170 words

A protein PP 21 , extracted from a source of said protein and having the following characteristics: (a) an electrophoretic mobility in the region of that of β 1 -globulins; (b) an isoelectric point of 4.6±0.3; (c) a sedimentation coefficient s 20 ,w of 3.2±0.2 S; (d) a molecular weight determined in an ultracentrifuge of 52,900±6,200; (e) an extinction coefficient E 1cm 1% (280 nm) of 10.5±1.0; (f) a carbohydrate fraction of 19.2±5.2 g/100 g (mannose 1.8±0.4, galactose 4.3±1.0, fucose 1.3±0.3, N-acetylglucosamine 6.5±2.0 and N-acetylneuraminic acid 5.3±1.5, each g/100 g); and (g) an amino acid composition as in the following table: ______________________________________ Coefficient Amino acid Residues per 100 residues of variation ______________________________________ lysine 8.75 3.91 histidine 1.58 6.81 arginine 2.75 6.07 aspartic acid 9.59 1.97 threonine 3.73 7.22 serine 7.43 5.95 glutamic acid 8.16 5.15 proline 11.34 9.36 glycine 7.01 1.25 alanine 4.98 3.62 cystine 1/2 5.50 0.84 valine 8.14 1.42 methionine 0.00 0.00 isoleucine 4.86 2.78 leucine 5.42 10.34 tyrosine 5.01 5.01 phenylalanine 3.56 1.00 tryptophan 2.12 17.65 ______________________________________

Claim 3

Dependent Claim 227 words

A process for obtaining or concentrating PP 21 as claimed in claim 1, which comprises subjecting an extract which has been obtained, using a dilute salt or buffer solution, from at least one organ which contains this protein to one or more of the following measures: (a) precipitation of the protein PP 21 using ammonium sulfate in the pH range 6 to 8 and at 30-60% saturation; (b) precipitation of accompanying proteins using an acridine base which is soluble in water, at a pH of 6 and a concentration of the base of 4 g/l or less, or precipitation of the protein PP 21 using an acridine base which is soluble in water, at a pH between 7 and 9 and a concentration of the base of 4 to 8 g/l; (c) precipitation of accompanying proteins by addition of up to 200 ml of ethanol per liter at pH 7; (d) preparative zone electrophoresis, the protein fraction having a mobility in the region of that of β1-globulins being obtained; (e) isoelectric focusing, the proteins in the pH range 4.4-4.8 being obtained; (f) gel filtration or ultrafiltration, the proteins in the molecular range of 40,000 to 100,000 being obtained; (g) adsorption onto a weakly basic ion exchanger, and elution of the protein PP 21 ; and (h) concentration by immunoadsorption, to obtain or concentrate said PP 21 protein.

Claim 4

Dependent Claim 21 words

The process of claim 2, wherein said organ is selected from the group consisting of placenta, stomach, intestines, bladder and spleen.

Claim 5

Dependent Claim 45 words

A process for isolating the protein PP 21 of claim 1 comprising the steps of subjecting a liquid containing said protein PP 21 to at least one known procedure for isolating proteins, and, in each instance, recovering that material containing said protein PP 21 .

Claim 6

Dependent Claim 18 words

The protein of claim 1, said protein being extracted from an organ or an extract of an organ.

Claim 7

Dependent Claim 20 words

The protein PP 21 of claim 5, said protein being extracted from a placenta or an extract of a placenta.

Claim 8

Dependent Claim 12 words

The protein PP 21 of claim 6, said protein being substantially pure.

Claim 9

Dependent Claim 14 words

The protein PP 21 of claim 1, wherein said protein is isolated and concentrated.

Claim 10

Dependent Claim 22 words

An antiserum to the protein of claim 1 obtained by immunizing an animal with the protein PP 21 and recovering said antiserum.

Claim 11

Dependent Claim 23 words

A method for detecting and determining protein PP 21 comprising the step of utilizing an effective amount of the antiserum of claim 9.

Claim 12

Dependent Claim 164 words

A substantially pure protein PP 21 , with the following characteristics: (a) an electrophoretic mobility in the region of that of β 1 -globulins; (b) an isoelectric point of 4.6±0.3; (c) a sedimentation coefficient s 20 ,w of 3.2±0.2 S; (d) a molecular weight determined in an ultracentrifuge of 52,900±6,200; (e) an extinction coefficient E 1cm 1% (280 nm) of 10.5±1.0; (f) a carbohydrate fraction of 19.2±5.2 g/100 g (mannose 1.8±0.4, galactose 4.3±1.0, fucose 1.3±0.3, N-acetylglucosamine 6.5±2.0 and N-acetylneuraminic acid 5.3±1.5, each g/100 g); and (g) an amino acid composition as in the following table: ______________________________________ Coefficient Amino acid Residues per 100 residues of variation ______________________________________ lysine 8.75 3.91 histidine 1.58 6.81 arginine 2.75 6.07 aspartic acid 9.59 1.97 threonine 3.73 7.22 serine 7.43 5.95 glutamic acid 8.16 5.15 proline 11.34 9.36 glycine 7.01 1.25 alanine 4.98 3.62 cystine 1/2 5.50 0.84 valine 8.14 1.42 methionine 0.00 0.00 isoleucine 4.86 2.78 leucine 5.42 10.34 tyrosine 5.01 5.01 phenylalanine 3.56 1.00 tryptophan 2.12 17.65 ______________________________________

Claim 13

Dependent Claim 164 words

A substantially pure protein PP 21 , with the following characteristics: (a) an electrophoretic mobility in the region of that of β 1 -globulins; (b) an isoelectric point of 4.6±0.3; (c) a sedimentation coefficient s 20 ,w of 3.2±0.2 S; (d) a molecular weight determined in an ultracentrifuge of 52,900±6,200; (e) an extinction coefficient E 1cm 1% (280 nm) of 10.5±1.0; (f) a carbohydrate fraction of 19.2±5.2 g/100 g (mannose 1.8±0.4, galactose 4.3±1.0, fucose 1.3±0.3, N-acetylglucosamine 6.5±2.0 and N-acetylneuraminic acid 5.3±1.5, each g/100 g); and (g) an amino acid composition as in the following table: ______________________________________ Coefficient Amino acid Residues per 100 residues of variation ______________________________________ lysine 8.75 3.91 histidine 1.58 6.81 arginine 2.75 6.07 aspartic acid 9.59 1.97 threonine 3.73 7.22 serine 7.43 5.95 glutamic acid 8.16 5.15 proline 11.34 9.36 glycine 7.01 1.25 alanine 4.98 3.62 cystine 1/2 5.50 0.84 valine 8.14 1.42 methionine 0.00 0.00 isoleucine 4.86 2.78 leucine 5.42 10.34 tyrosine 5.01 5.01 phenylalanine 3.56 1.00 tryptophan 2.12 17.65 ______________________________________

Claim 14

Dependent Claim 55 words

A method for diagnosing and monitoring a disease or monitoring the treatment of said disease, said disease being accompanied by an increase in the concentration of protein PP 21 comprising the step of utilizing in an immunochemical method the protein PP 21 of claim 1 to achieve said intended purpose with respect to said disease.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	1989-06-09	1989-12-09	1990-06-10	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	1993-06-09	1993-12-09	1994-06-10	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	1997-06-09	1997-12-09	1998-06-10	$7700.00	Window Closed / Lapsed

Tissue protein PP21, a process for obtaining it and its use

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